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TIMP3 interplays with apelin to regulate cardiovascular metabolism in hypercholesterolemic mice.

Stöhr R, Kappel BA, Carnevale D, Cavalera M, Mavilio M, Arisi I, Fardella V, Cifelli G, Casagrande V, Rizza S, Cattaneo A, Mauriello A, Menghini R, Lembo G, Federici M - Mol Metab (2015)

Bottom Line: ApoE(-/-)TIMP3(-/-) revealed decreased lifespan.Metabolomics analysis revealed an increase in circulating markers of oxidative stress with a reduction in long chain fatty acids.Apelin is a regulator of fatty acid oxidation, and we found a reduction in the levels of enzymes involved in fatty acid oxidation in the left ventricle of ApoE(-/-)TIMP3(-/-) mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, Italy ; Department of Internal Medicine I, University Hospital Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.

ABSTRACT

Objective: Tissue inhibitor of metalloproteinase 3 (TIMP3) is an extracellular matrix (ECM) bound protein, which has been shown to be downregulated in human subjects and experimental models with cardiometabolic disorders, including type 2 diabetes mellitus, hypertension and atherosclerosis. The aim of this study was to investigate the effects of TIMP3 on cardiac energy homeostasis during increased metabolic stress conditions.

Methods: ApoE(-/-)TIMP3(-/-) and ApoE(-/-) mice on a C57BL/6 background were subjected to telemetric ECG analysis and experimental myocardial infarction as models of cardiac stress induction. We used Western blot, qRT-PCR, histology, metabolomics, RNA-sequencing and in vivo phenotypical analysis to investigate the molecular mechanisms of altered cardiac energy metabolism.

Results: ApoE(-/-)TIMP3(-/-) revealed decreased lifespan. Telemetric ECG analysis showed increased arrhythmic episodes, and experimental myocardial infarction by left anterior descending artery (LAD) ligation resulted in increased peri-operative mortality together with increased scar formation, ventricular dilatation and a reduction of cardiac function after 4 weeks in the few survivors. Hearts of ApoE(-/-)TIMP3(-/-) exhibited accumulation of neutral lipids when fed a chow diet, which was exacerbated by a high fat, high cholesterol diet. Metabolomics analysis revealed an increase in circulating markers of oxidative stress with a reduction in long chain fatty acids. Using whole heart mRNA sequencing, we identified apelin as a putative modulator of these metabolic defects. Apelin is a regulator of fatty acid oxidation, and we found a reduction in the levels of enzymes involved in fatty acid oxidation in the left ventricle of ApoE(-/-)TIMP3(-/-) mice. Injection of apelin restored the hitherto identified metabolic defects of lipid oxidation.

Conclusion: TIMP3 regulates lipid metabolism as well as oxidative stress response via apelin. These findings therefore suggest that TIMP3 maintains metabolic flexibility in the heart, particularly during episodes of increased cardiac stress.

No MeSH data available.


Related in: MedlinePlus

Loss of TIMP3 leads to disrupted apelin secretion from heart while supplementation can correct some of the metabolic abnormalities in the heart. A) Apelin expression by qPCR in heart, muscle, adipose tissue and liver reveals a difference between ApoE−/−TIMP3−/− and ApoE−/− animals only in the heart (*p < 0.05, one-way ANOVA, n = 5 per group). B) Circulating apelin levels measured by ELISA were also found to be reduced in the ApoE−/−TIMP3−/− (*p < 0.05, Student's t test, n = 4 per group). C) Single dose of intraperitoneally injected apelin restores impaired lipid oxidation of ApoE−/−TIMP3−/− mice during the prolonged fasting state (interval of 8–12 h) measured by indirect calorimetry. Data are mean ± SEM (****p < 0.001, one-way ANOVA, n = 5–6 per group).
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fig7: Loss of TIMP3 leads to disrupted apelin secretion from heart while supplementation can correct some of the metabolic abnormalities in the heart. A) Apelin expression by qPCR in heart, muscle, adipose tissue and liver reveals a difference between ApoE−/−TIMP3−/− and ApoE−/− animals only in the heart (*p < 0.05, one-way ANOVA, n = 5 per group). B) Circulating apelin levels measured by ELISA were also found to be reduced in the ApoE−/−TIMP3−/− (*p < 0.05, Student's t test, n = 4 per group). C) Single dose of intraperitoneally injected apelin restores impaired lipid oxidation of ApoE−/−TIMP3−/− mice during the prolonged fasting state (interval of 8–12 h) measured by indirect calorimetry. Data are mean ± SEM (****p < 0.001, one-way ANOVA, n = 5–6 per group).

Mentions: To gain insight into the regulation of glucose and lipid metabolism of the heart by TIMP3 we performed mRNA transcriptomic analysis by Next Generation Sequencing (Illumina) of the heart in ApoE−/−TIMP3−/− and ApoE−/− littermates. We identified several significantly upregulated and downregulated transcripts from ApoE−/−TIMP3−/− compared with ApoE−/− littermates (Supplementary Figure S4A and Supplementary Table S4). Among the downregulated transcripts, confirmed after qPCR, we found apelin (Supplementary Figure S4A,B), which we confirmed to be significantly reduced in heart tissue but not in the epididymal White Adipose Tissue (eWAT), muscle and liver from ApoE−/−TIMP3−/− mice by quantitative PCR (*p < 0.05, Student's t test, Figure 7A). Apelin blood levels measured by ELISA were shown to be significantly lower in the ApoE−/−TIMP3−/− compared with ApoE−/− littermates (*p < 0.05, Student's t test, Figure 7B).


TIMP3 interplays with apelin to regulate cardiovascular metabolism in hypercholesterolemic mice.

Stöhr R, Kappel BA, Carnevale D, Cavalera M, Mavilio M, Arisi I, Fardella V, Cifelli G, Casagrande V, Rizza S, Cattaneo A, Mauriello A, Menghini R, Lembo G, Federici M - Mol Metab (2015)

Loss of TIMP3 leads to disrupted apelin secretion from heart while supplementation can correct some of the metabolic abnormalities in the heart. A) Apelin expression by qPCR in heart, muscle, adipose tissue and liver reveals a difference between ApoE−/−TIMP3−/− and ApoE−/− animals only in the heart (*p < 0.05, one-way ANOVA, n = 5 per group). B) Circulating apelin levels measured by ELISA were also found to be reduced in the ApoE−/−TIMP3−/− (*p < 0.05, Student's t test, n = 4 per group). C) Single dose of intraperitoneally injected apelin restores impaired lipid oxidation of ApoE−/−TIMP3−/− mice during the prolonged fasting state (interval of 8–12 h) measured by indirect calorimetry. Data are mean ± SEM (****p < 0.001, one-way ANOVA, n = 5–6 per group).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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Show All Figures
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fig7: Loss of TIMP3 leads to disrupted apelin secretion from heart while supplementation can correct some of the metabolic abnormalities in the heart. A) Apelin expression by qPCR in heart, muscle, adipose tissue and liver reveals a difference between ApoE−/−TIMP3−/− and ApoE−/− animals only in the heart (*p < 0.05, one-way ANOVA, n = 5 per group). B) Circulating apelin levels measured by ELISA were also found to be reduced in the ApoE−/−TIMP3−/− (*p < 0.05, Student's t test, n = 4 per group). C) Single dose of intraperitoneally injected apelin restores impaired lipid oxidation of ApoE−/−TIMP3−/− mice during the prolonged fasting state (interval of 8–12 h) measured by indirect calorimetry. Data are mean ± SEM (****p < 0.001, one-way ANOVA, n = 5–6 per group).
Mentions: To gain insight into the regulation of glucose and lipid metabolism of the heart by TIMP3 we performed mRNA transcriptomic analysis by Next Generation Sequencing (Illumina) of the heart in ApoE−/−TIMP3−/− and ApoE−/− littermates. We identified several significantly upregulated and downregulated transcripts from ApoE−/−TIMP3−/− compared with ApoE−/− littermates (Supplementary Figure S4A and Supplementary Table S4). Among the downregulated transcripts, confirmed after qPCR, we found apelin (Supplementary Figure S4A,B), which we confirmed to be significantly reduced in heart tissue but not in the epididymal White Adipose Tissue (eWAT), muscle and liver from ApoE−/−TIMP3−/− mice by quantitative PCR (*p < 0.05, Student's t test, Figure 7A). Apelin blood levels measured by ELISA were shown to be significantly lower in the ApoE−/−TIMP3−/− compared with ApoE−/− littermates (*p < 0.05, Student's t test, Figure 7B).

Bottom Line: ApoE(-/-)TIMP3(-/-) revealed decreased lifespan.Metabolomics analysis revealed an increase in circulating markers of oxidative stress with a reduction in long chain fatty acids.Apelin is a regulator of fatty acid oxidation, and we found a reduction in the levels of enzymes involved in fatty acid oxidation in the left ventricle of ApoE(-/-)TIMP3(-/-) mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Systems Medicine, University of Rome Tor Vergata, 00133 Rome, Italy ; Department of Internal Medicine I, University Hospital Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.

ABSTRACT

Objective: Tissue inhibitor of metalloproteinase 3 (TIMP3) is an extracellular matrix (ECM) bound protein, which has been shown to be downregulated in human subjects and experimental models with cardiometabolic disorders, including type 2 diabetes mellitus, hypertension and atherosclerosis. The aim of this study was to investigate the effects of TIMP3 on cardiac energy homeostasis during increased metabolic stress conditions.

Methods: ApoE(-/-)TIMP3(-/-) and ApoE(-/-) mice on a C57BL/6 background were subjected to telemetric ECG analysis and experimental myocardial infarction as models of cardiac stress induction. We used Western blot, qRT-PCR, histology, metabolomics, RNA-sequencing and in vivo phenotypical analysis to investigate the molecular mechanisms of altered cardiac energy metabolism.

Results: ApoE(-/-)TIMP3(-/-) revealed decreased lifespan. Telemetric ECG analysis showed increased arrhythmic episodes, and experimental myocardial infarction by left anterior descending artery (LAD) ligation resulted in increased peri-operative mortality together with increased scar formation, ventricular dilatation and a reduction of cardiac function after 4 weeks in the few survivors. Hearts of ApoE(-/-)TIMP3(-/-) exhibited accumulation of neutral lipids when fed a chow diet, which was exacerbated by a high fat, high cholesterol diet. Metabolomics analysis revealed an increase in circulating markers of oxidative stress with a reduction in long chain fatty acids. Using whole heart mRNA sequencing, we identified apelin as a putative modulator of these metabolic defects. Apelin is a regulator of fatty acid oxidation, and we found a reduction in the levels of enzymes involved in fatty acid oxidation in the left ventricle of ApoE(-/-)TIMP3(-/-) mice. Injection of apelin restored the hitherto identified metabolic defects of lipid oxidation.

Conclusion: TIMP3 regulates lipid metabolism as well as oxidative stress response via apelin. These findings therefore suggest that TIMP3 maintains metabolic flexibility in the heart, particularly during episodes of increased cardiac stress.

No MeSH data available.


Related in: MedlinePlus